Department of Anesthesiology, E-DA Hospital/I-Shou University, Kaohsiung 824, Taiwan, R.O.C.
Oncol Rep. 2013 Nov;30(5):2304-10. doi: 10.3892/or.2013.2722. Epub 2013 Sep 5.
Propofol is one of the most widely clinically used intravenous anesthetic, and it induces apoptosis in human and murine leukemia cell lines. Yet, whether propofol causes DNA damage and affects the mRNA expression of repair-associated genes in cancer cells remains undetermined. In the present study, we investigated the effects of propofol on DNA damage and associated mRNA gene expression in RAW264.7 cells. Comet assay and DNA gel electrophoresis were used to evaluate DNA damage in RAW264.7 cells and propofol-inhibited cell growth in vitro. The results revealed a longer DNA tail and DNA fragmentation. Real-time PCR assay was used to examine mRNA gene expression of DNA damage and DNA repair-associated genes. Following exposure to propofol for 48 h, a decrease in the mRNA expression of DNA-PK, BRCA1, MGMT and p53 was noted in the RAW264.7 cells. Results from the western blotting indicated that p53, MGMT, 14-3-3-σ, BRCA1 and MDC1 proteins were decreased while p-p53 and p-H2A.X(S140) were increased in the RAW264.7 cells following exposure to propofol. In conclusion, exposure to propofol caused DNA damage and inhibited mRNA expression and protein levels of repair-associated genes in RAW264.7 cells.
异丙酚是临床上最广泛使用的静脉麻醉剂之一，它可诱导人源和鼠源白血病细胞系凋亡。然而，异丙酚是否会导致 DNA 损伤，并影响癌细胞中与修复相关的基因的 mRNA 表达，目前仍不清楚。在本研究中，我们研究了异丙酚对 RAW264.7 细胞中 DNA 损伤和相关 mRNA 基因表达的影响。彗星试验和 DNA 凝胶电泳用于评估 RAW264.7 细胞中的 DNA 损伤和异丙酚抑制细胞体外生长的情况。结果显示 DNA 尾巴更长，发生了 DNA 片段化。实时 PCR 检测用于检测与 DNA 损伤和 DNA 修复相关基因的 mRNA 基因表达。暴露于异丙酚 48 小时后，RAW264.7 细胞中 DNA-PK、BRCA1、MGMT 和 p53 的 mRNA 表达减少。Western blot 结果表明，暴露于异丙酚后，RAW264.7 细胞中的 p53、MGMT、14-3-3-σ、BRCA1 和 MDC1 蛋白减少，而 p-p53 和 p-H2A.X(S140)增加。综上所述，暴露于异丙酚可导致 RAW264.7 细胞的 DNA 损伤，并抑制与修复相关的基因的 mRNA 表达和蛋白水平。
